PROJECT SUMMARY Healing of chronic wounds is impaired by the presence of necrotic tissue, which contains dead cells and debris within second (deep partial thickness) and third degree (full thickness) burns (eschar) and chronic ulcers (e.g., diabetic ulcers). The accumulation of non-viable tissue in a chronic wound can result in a prolonged inflammatory response and mechanical obstruction of the wound contraction process with impeded granulation and reepithelialization, preventing wounds from healing. In addition, necrotic tissue is susceptible to bacterial infection, further impeding wound healing, and potentially inducing sepsis in severe cases. Debridement of necrotic tissue is a basic requirement needed to facilitate wound closure. To remove necrotic tissue from wounds, various types of debridement techniques are utilized, including surgical debridement, enzymatic debridement, autolytic debridement, biological debridement, and mechanical debridement, with surgical and enzymatic debridement being most prevalent. Although surgical debridement is the fastest and the most efficient method for debriding necrotic tissue, this is performed by trained doctors/surgeons (no nursing or clinical staff). Viable tissue is inevitably also debrided because of a lack of a clear border between necrotic tissue and viable tissue, resulting in an enlarged wound area and increased blood loss. Enzymatic debridement, while slower, requires less training and is generally performed by nursing and other clinical staff. There is a clinical need for a debridement agent that can efficiently remove necrotic tissue without loss of viable tissue, is easy to use, and can control infection. The overall goal of this project is to develop an enzymatic debriding product that can fully debride necrotic tissue faster than existing products, is easy to apply and to remove, and is more compatible with medically compromised senior citizens. An innovative enzyme- based debridement system has been developed to fulfill these requirements. The first specific aim is to conduct biocompatibility testing for product safety of the enzymatic debrider by in vitro and in vivo testing. The second specific aim is to conduct preclinical in vivo testing for debrider efficacy utilizing pig full thickness burn eschar and acid-induced necrosis. The third specific aim is to initiate product stability and shelf-life testing for at least one year. The fourth specific aim is to prepare documentation for human ?proof of efficacy? testing and for regulatory submission. Successful development of this debridement formulation will fulfill a number of unmet needs of current debridement procedures. The final product is to achieve effective debridement of necrotic tissue while being easily utilized, increasing treatment time efficiency, reducing medical costs, and improving wound healing outcomes.